PHYTOCHEMICAL PROFILES, ANTIBACTERIAL and ANTIFUNGAL ACTIVITY of LEAVES from the Psydrax Dicoccos (Gaertn) D

Available online at www.jpsscientificpublications.com Volume – 2; Issue - 1; Year – 2016; Page: 443 – 452

Indo – Asian Journal of Multidisciplinary Research (IAJMR)

ISSN: 2454-1370

PHYTOCHEMICAL PROFILES, ANTIBACTERIAL AND ANTIFUNGAL ACTIVITY OF LEAVES FROM THE Psydrax dicoccos (Gaertn) D. Umaiyambigai*, K. Saravanakumar and G. Adaikala Raj, Department of Botany, Annamalai University, Annamalai Nagar – 608 002, Tamil Nadu, India. Abstract The phytochemical analyses and antimicrobial activity of Psydrax dicoccos leaves were extracted successively with different solvents viz., petroleum ether, chloroform, ethyl acetate and methanol and screened for their antimicrobial activity against Staphylococcus aureus, Streptococcus pyogenes, Enterococcus faecalis, Escherichia coli, Proteus vulgaris, Pseudomonas aeruginosa, Vibrio cholerae, Candida albicans, C. parapsilosis and C. tropicalis by using Disc diffusion method. Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC) and Minimum Fungicidal Concentration (MFC) were also determined in the present research. The methanol extract of Psydrax dicoccos showed the highest antimicrobial activity against all the bacterial and fungal strains tested than the other extracts. The mean zones of inhibition produced by the extracts in Agar diffusion assays against the tested bacterial strains was ranged from 7.0 to 22.5 mm. The MIC values were between 125 and 250 μg/ml, MBC values 250 and 1000 μg/ml and MFC 500 and 1000 μg/ml values were recorded. Phytochemical analyses of different extracts of Psydrax dicoccos leaves was analyzed. The methanol extract of Psydrax dicoccos leaves showed the presence of strong phytochemicals viz., steroids, tannins, phenolic compounds and terpenoids compared to other extracts. The highest mean of zone inhibition (22.5 mm) was observed in the methanol extract of Psydrax dicoccos against the bacteria Staphylococcus aureus. From the above findings, we concluded that the methanol extract of Psydrax dicoccos will be needed to isolate and characterize the compounds. Key words: Antimicrobial activity, Psydrax dicoccos, MIC, MBC and MFC.

1. Introduction Psydrax dicoccos belongs to the family was used externally for rheumatic pains (Neelima Rubiaceae and commonly called Ceylon boxwood et al., 2011). and Nazhuvai, Irambarathaan in Tamil and Microbial infections are major public distributed in all parts of tropical and sub-tropical health problems in the developed countries. region of India. All parts of the plant have been Antibiotics are used to treat these infections. Due recognized to have medicinal properties. Plant to indiscriminate use of commercial antibiotics, possesses antipyretic activity. In India, bark was the incidence of multiple antibiotic resistances in used as febrifuge and also applied as plasters. The human pathogens is increasing. This has forced decoction of roots was used internally for treating the scientists to search for new antimicrobial diarrhoea. Bark powder boiled with sesame oil substances from various sources like medicinal plants. Medicinal plants constitute the main source of new pharmaceuticals and health care products *Corresponding author: D. Umaiyambigai (Ivanona et al., 2005). The use of traditional E. mail: [email protected] medicines was widespread in India (Jeyachandran, Received: 10.12.2015; Revised: 17.12.2015; Accepted: 25.12.2015. et al., 2007).

Infectious disease caused by bacteria, have become one of the great challenges for viruses, fungi and parasites are still a major threat modern healthcare. Majority of scientists define to public health, despite the tremendous progress MDR as ˝resistance to at least 3 classes of in human medicine (Cosa, 2006). The past three antimicrobial agents˝ (Falagas et al., 2006). decades have seen a dramatic increase in Antimicrobial agents have substantially reduced microbial resistance to antimicrobial agents the threat posed by infectious diseases over a (Chopra, 1996). Such situation stimulates the period of time since their discovery in the 1940’s development of new anti-microbial agents in order (Lewis and Ausubel, 2006). However, the to treat the infectious disease in an effective escalation of multidrug resistance in bacteria in manner. So, this matter continued to an era to recent years has seriously jeopardized these gains. identify the potential antimicrobial agent from the This has gained worldwide attention due to the natural resources. The edible plants that used for high impact on public health. Increased usage of traditional medicine contain a wide range of antimicrobial agents to treat bacterial infections substance that can be used to treat abundant of has lead to the emergence of MDR strains infectious disease with reduced side effects (Bonnet, 2004). Invasive fungal infections are of (Duraipandiyan, 2006). In Asia, the use of great concern for human beings because they are medicinal plants to cure specific illness has been associated with unacceptable high mortality rates. use for many years (Bhattacharjee, 1998). More than 90 % of all reported fungal - related Furthermore, Malaysia is rich in various edible deaths result from species that belong to one of plants with diverse biological & pharmacological three genera: Cryptococcus, Candida and properties (Yoga, 2005). Aspergillus. In turn, superficial infections of the skin and nails are the most common fungal The numbers of resistant strains of microbial pathogens are growing since penicillin diseases in humans, affecting C. albicans 25 % of resistant and multiresistant Pneumococci caused a the population worldwide (Karan, 2009). Hence, major problem in South African hospitals in 1977. present study was carried out to evaluate the (Berkowitz, 1995) calls the emerging of drug phytochemical properties and antimicrobial resistant bacteria a medical catastrophe. Leggiadro activity of petroleum ether, chloroform, ethyl (1995) stated that effective regimens may not be acetate and methanol extracts of leaves of Psydrax available to treat some Enterococcal isolates and dicoccos against bacterial and fungal strains. that it is critically important to develop new 2. Materials and methods antimicrobial compounds for these and other Collection of Plant material and preparation of organisms before we enter the post-antibiotic era. extracts New compounds inhibiting microorganisms such as benzoin and emetine have been isolated from The fresh leaves of Psydrax dicoccos plants (Cox, 1994). The antimicrobial compounds were collected from Authukurichi region from plants may inhibit bacteria by a different (11°24′37″N 79°21′4″E), Ariyalur District, Tamil mechanism than the presently used antibiotics and Nadu, India. During the months from March to April may have clinical value in treatment of resistant 2014, the specimens were deposited in the Herbarium microbial strains. A number of traditional natural of Department of Botany, Annamalai University, products have been increased and much work has Annamalai Nagar. Collected leaves were initially been done on selected ethno medicinal plants for washed with water, then surface sterilized with antibacterial activity against pathogenic strains of disinfectant solution of 10 % sodium hypochlorite Gram negative and Gram positive bacteria (Singh, solution and finally rinsed with sterile distilled 2002). water and shade dried under room temperature and grounded in to a coarse powder. One hundred Bacterial infections especially those grams of coarse powder was extracted with caused by multi-drug resistant (MDR) bacteria different organic solvents like non-polar to polar

Umaiyambigai/ Indo – Asian Journal of Multidisciplinary Research (IAJMR), 2(1): 443 – 452 445 viz., Petroleum ether, chloroform, ethyl acetate containing 106 CFU/ml were swabbed on the top and methanol for 8 hours using Soxhlet apparatus of the solidified media and the plates were dried and the solvents were evaporated under vacuum in and uniformly spread. After drying, the extracts a rotary evaporator (Heidolph, Germany) and the were placed on the disc with sterile forceps and dried powder was stored at 4˚C for further use. gently pressed to ensure the contact with the incubated agar surface. Ciprofloxacin (5 µg/disc) Phytochemical analysis The solvents such as, petroleum ether, for bacteria and Amphotercin-B (100 units/disc) for chloroform, ethyl acetate and methanol extracts of yeast were used as positive control. Ten per cent leaves of Psydrax dicoccos were used for DMSO was used as blind control. Finally, the qualitative phytochemical analyses. Various inoculated plates were incubated at 37 °C for 24 bioactive phytochemicals namely, flavonoids, hrs. The zone of inhibition was observed and tannins, steroids, glycosides, saponins, phenolic measured in millimeters. Each experiment was compounds, terpenoids and alkaloids were carried out in triplicates. analyzed according to described by Harborne Minimum Inhibitory Concentration (MIC) (1983) and Trease (1983). Minimum inhibitory concentrations of plant different extracts were tested in MHB for bacteria Microorganisms The seven clinical bacterial isolates and and SDB for fungi described by Ericsson and three fungal species were obtained from Raja Sherris (2002). The extracts were dissolved in 10 % Muthaiah Medical College Hospital, Annamalai DMSO to obtain 2 mg/ml and 0.5 ml of stock University, Tamil Nadu. Gram positive bacteria: solution was incorporated into 0.5 ml of MHA to get Staphylococcus aureus, Streptococcus pyogenes, concentrations of 1000, 500, 250, 125, 62.5, 31.2 Enterococcus faecalis and Gram negative bacteria: and 15.6 µg/ml, 100 µl as standard. Bacterial Escherichia coli, Proteus vulgaris, Pseudomonas suspension of the test organism was transferred into aeruginosa, Vibrio cholerae and fungal species, each tube and incubated at 37 °C for 24 hrs. The Candida albicans, Candida parapsilosis and tube devoid of plant extracts kept as control and the Candida tropicalis were tested in the present MIC of the extracts were examined. study. The stock cultures were maintained on Minimum Bactericidal Concentration (MBC) Muller Hinton Agar medium and Sabouraud’s The MBC of the different extracts were Dextrose Agar at 4 °C separately for both bacteria determined by plating 100 µl of samples from and fungus respectively. In vitro antibacterial each MIC assay tube with growth inhibition into activity of the isolates was determined by using freshly prepared MHB and the plates were Muller Hinton Agar (MHA) and Muller Hinton incubated at 37 °C for 24 hrs bacteria. The MBC Broth (MHB). In vitro antifungal activity was values were recorded as the lowest concentration determined by using Sabouraud’s Dextrose Agar of the extracts that did not permit any visible (SDA) and Sabouraud’s Dextrose Broth (SDB) bacterial colony growth on the agar plate during respectively. the period of incubation. Antibacterial and Antifungal assays Minimum Fungicidal Concentration (MFC) The MFC of the different extracts were Disc diffusion method The agar diffusion method (Bauer et al., determined by plating 100 µl of samples from 1966) was employed for the initial assessment of each MIC assay tube with growth inhibition into antimicrobial potential of the extracts. Sterilized freshly prepared SDA and the plates were mediums of 20 ml MHA (Bacteria) and 20 ml incubated at 37 °C for 24 hrs yeast. The MFC SDA (Fungi) were separately poured in each petri values were recorded as the lowest concentration of the extracts that did not permit any visible plate and kept untouched until it solidify. The standard inoculum of bacterial suspension bacterial colony growth on the agar plate during containing108 CFU/ml and suspension of yeast the period of incubation.

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Table - 1: Phytochemical analysis of the different extracts of leaves Psydrax dicoccos

S.No Phytoconstituents Petroleum ether Chloroform Ethyl acetate Methanol

1 Alkaloids + + + + 2 Flavonoids - - - + 3 Phenolic compounds - - + ++ 4 Saponins - - - + 5 Tannins - - + ++ 6 Cardiac glycosides + + + + 7 Steroids + + + + + 8 Terpenoids - + - + + (++) = Strong; (+) = Positive (present); (-) =Negative (absent) 3. Results The petroleum ether, chloroform, ethyl petroleum ether, chloroform, ethyl acetate and acetate and methanol extracts of the leaf of methanol using Soxhlet apparatus. The extracts were Psydrax dicoccos revealed that the presence of evaluated for antibacterial activities against Gram phytochemicals such as alkaloids, flavonoids, positive clinical isolates viz., Staphylococcus aureus cardiac glycosides, phenolic compounds, Streptococcus pyogenes and Enterococcus faecalis, saponins, steroids, tannins and terpenoids. The Gram negative clinical isolates viz., Escherichia coli, methanol extracts of Psydrax dicoccos revealed Pseudomonas aeruginosa, Proteus vulgaris and the presence of strong phytochemicals, terpenoids, Vibrio cholerae and antifungal activities against saponins, steroids, flavonoids and phenolic Candida albicans, Candida parapsilosis and compounds. The ethyl acetate extracts were present Candida tropicalis. in all phytochemicals except cardiac glycosides and The results clearly demonstrated that the saponins. The chloroform extracts were present in all methanol extract of Psydrax dicoccos leaves were the phytochemical except flavonoids, saponins, exhibited broad spectrum antimicrobial activity. The steroids, and tannins. The petroleum either extracts results of antimicrobial activity of different were absent in all the phytochemical except alkaloids, concentrations of petroleum ether, chloroform, ethyl flavonoids, saponins, and cardiac glycosides. acetate and methanol extracts of Psydrax dicoccos The plant extracts were prepared from leaves against bacterial and fungal strains. different parts (leaves) of Psydrax dicoccos by using the solvents with different polarity such as

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Table – 2: Antimicrobial activity of different extracts of leaves of Psydrax dicoccos

Mean zone of inhibitiona (mm)b Microbial strains Concentration of the extracts (g/disc) MIC MBC S. No. Ciprofloxacin (5 (g/mL) (g/mL) 1000 500 250 g/disc) 1 Staphylococcus aureus Petroleum ether 13.5± 0.50 10.8 ±0.76 9.3 ±0.57 29.1 ± 057 250 500 Chloroform 15.8± 0.67 13.1±0.28 10.8± 0.76 28.6± 0.76 250 500

Ethyl acetate 20.1±0.28 16.1± 0.28 12.3± 0.57 29.0 ± 0.50 125 250 Methanol 22.5± 0.50 17.3 ±0.57 13.0± 0.50 30.1± 0.78 125 250 2 Streptococcus pyogenes Petroleum ether 13.1 ±0.28 9.8 ± 0.76 8.1± 0.28 30.0 ± 0.28 250 500 Chloroform 14.6± 0.76 10.3± 0.76 8..5± 0.50 29.6 ± 0.75 250 500

Ethyl acetate 17.8± 0.76 12.5± 0.50 10.5± 0.50 28.3± 0.50 125 250 Methanol 18.5±0.50 14.1± 0.28 11.8 ± 0.57 28.1± 0.28 125 250 3 Enterococcus faecalis Petroleum ether 12.0± 0.50 9.5 ± 0.50 7.8 ± 0.76 26.7± 0.76 500 1000 Chloroform 12.6± 0.50 10.1± 0.28 9.1 ± 0.28 29.0 ± 0.50 500 1000

Ethyl acetate 13..6 ± 0.36 10.5± 0.50 9.6± 0.76 28.1± 0.28 250 500 Methanol 14.8± 0.76 11.1 ± 0.78 10.1 ± 0.28 29.3 ± 0.50 250 500 4 Escherichia coli Petroleum ether 11.6 ± 0.67 9.3± 0.57 7.5 ± 0.50 30.0 ± 0.57 500 1000 Chloroform 12.1± 0.78 9.8± 0.76 8.0 ± 0.50 29.8 ± 0.76 500 1000

Ethyl acetate 13.1± 0.28 10.0 ± 0.70 9.1 ± 0.28 28.5± 0.50 250 500 Methanol 13.5± 0.50 10.3 ± 0.57 9.5± 0.50 30.1± 0.28 250 500 5 Proteus vulgaris Petroleum ether 10.3± 0.57 9.8± 0.76 7.1 ± 0.28 26.5 ± 0.28 500 1000 Chloroform 10.6 ± 0.56 9.0 ± 0.50 7.8± 0.76 29.1 ± 0.28 500 1000

Ethyl acetate 11.3± 0.57 9.8± 0.67 8.6± 0.76 30.5± 0.50 250 500 Methanol 13.1 ± 0.35 10.5± 0.50 9.3 ± 0.57 27.1 ± 0.28 250 500 aDiameter of zone of inhibition (mm) including the disc diameter of 6 mm bMean of three assays; ± - Standard deviation ; Significant at P<0.05

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Mean zone of inhibitiona (mm)b

Concentration of the extracts (g/disc) MIC MBC/MFC S. No. Microbial strains (g/mL) (g/mL) Ciprofloxacin 1000 500 250 (10g/disc)

6 Pseudomonas aeruginosa Petroleum ether 10.8± 0.76 9.1± 0.28 7.3 ± 0.57 29.8 ± 0.76 500 1000 Chloroform 11.3± 0.55 9.8± 0.76 7.8± 0.76 26.7± 0.58 500 1000

Ethyl acetate 12.3 ± 0.57 10.1 ± 0.28 8.6 ± 0.76 28.0 ± 0.50 250 500 Methanol 13.0± 0.50 10.3± 0.75 9.5 ± 0.50 30.1± 0.28 250 500 7 Vibrio cholera Petroleum ether 11.0 ± 0.50 8.5 ± 0.50 7.3 ± 0.57 21.1 ± 0.28 500 1000 12.8± 0.76 9.8± 0.76 8..1± 0.28 26.3 ± Chloroform 500 1000 0.57 Ethyl acetate 12.1 ± 0.28 10.8 ± 0.76 9.3± 0.57 26.0 ± 0.50 250 500 Methanol 13.8 ± 0.76 11.1± 0.28 9.6± 0.76 30.0 ± 0.50 250 500 8 Candida albicans Petroleum ether 10.5± 0.50 9.1 ± 0.50 7.1 ± 0.28 15.1 ± 0.28 500 1000 Chloroform 11..0± 0.50 9.1± 0.50 7.8± 0.76 13.5± 0.50 500 1000

Ethyl acetate 12.8 ± 0.76 11.0 ± 0.50 8.5± 0.57 11.0± 0.50 250 500 Methanol 13.8 ± 0.50 11.8 ± 072 9.1± 0.28 15.3± 0.57 250 500 9 Candida parapsilosis Petroleum ether 10.0 ± 0.50 8.1 ± 0.78 7.0± 0.50 16.3 ± 0.57 500 1000 Chloroform 11..5 ±0.50 8.3±0.50 7.5± 0.50 14.1± 0.28 500 1000

Ethyl acetate 13.0 ± 0.50 11.1± 0.28 7.8 ± 0.76 15.3± 0.57 250 500 Methanol 14.5± 0.50 1!.8± 0.28 9.3 ± 0.57 14.0± 0.50 250 500 10 Candida tropicalis Petroleum ether 9.0± 0.50 8.1 ± 0.28 7.0 ± 0.50 16.0 ± 0.50 500 1000 Chloroform 10.1± 0.28 8.1± 0.28 7.5± 0.50 14.7 ± 0.58 500 1000

Ethyl acetate 11.5 ± 0.50 9.5± 0.50 8.1 ± 0.28 16.3 ± 0.57 250 500 Methanol 12.8 ± 0.76 10.1± 0.78 9.0± 0.50 15.0± 0.50 250 500 aDiameter of zone of inhibition (mm) including the disc diameter of 6 mm bMean of three assays; ± - Standard deviation ; Significant at P<0.05

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The results revealed that the highest mean extract of Psydrax dicoccos showed the highest zone of inhibition produced by the leaf extracts of antibacterial activity against S. aureus followed by Psydrax dicoccos for bacteria ranged between 7.1 other strains. The highest mean zone of inhibition  0.28 and 22.5  0.50 mm. For fungi, the mean (22.5 mm) and the lowest MIC value (125 µg/ml) zone of inhibition produced by the leaf and MBC value (250 µg/ml) were observed in extracts ranged between 7.0  0.50 and 14.5  methanol leaf extract. In antifungal activity, the 0.50 mm. The MIC and MBC values of crude methanol leaf extract of Psydrax dicoccos showed extracts of leaves of Psydrax dicoccos for highest antifungal activity and it was followed by bacteria were between 62.5 and 500 µg/mL petroleum ether, chloroform and ethyl acetate. and 125 and 1000 µg/mL respectively. Similar results were also observed in The MIC and MFC values of crude extracts previous studies, methanol extracts have been of leaves of Psydrax dicoccos for fungi were shown to result in high extraction yields with between 250 and 500 µg/mL and 500 and 1000 strong antibacterial activities (Jo et al., 2012). µg/mL respectively. The highest mean zone Prashanth et al. (2001) proved that methanolic of inhibition for bacteria (22.5 ± 0.50 mm at extracts of pomegranate peels were more active 1000 µg/disc), lowest MIC (62.5 µg/mL) and than water extracts against Escherichia coli, MFC (250 µg/mL) values were recorded in Staphylococcus aureus and Bacillus subtilis. methanol extract of leaves of Psydrax dicoccos Ashik Mosaddik et al. (2000) reported that the against Staphylococcus aureus. The highest mean zone methanol extracts of Alangium salviifolium of inhibition for fungi (14.5 ± 0.50 mm at 1000 flowers showed a wide spectrum of antibacterial µg/disc), lowest MIC (250 µg/mL) and MFC (500 activity against Bacillus subtilis, Bacillus µg/mL) values were recorded in methanol extracts of megaterium and Staphylococcus aureus and Gram leaves of Psydrax dicoccos against Candida negative bacteria viz., Escherichia coli, Shigella parapsilosis and Candida albicans. sonnei, Shigella shiga, Shigella boydii, Salmonella typhi and Klebsiella sp. Ciprofloxacin (5 µg/disc) was used as the positive control for bacteria and the mean zones of The methanol extract of Psydrax dicoccos inhibitions were between 26.7  0.76 mm leaves showed the presence of strong and 31.5  0.50 mm. Amphotericin-B (100 phytochemicals viz., steroids, tannins, phenolic units/disc) was used as the positive controls for compounds and terpenoids compared to other Candida species and the mean zones of inhibition extracts. were between 11.0  0.50 mm and 16.3  0.57 Methanol extracts of Psydrax dicoccos mm. For bacteria, the lowest activity (7.1  showed potential inhibitory action on tested fungal 0.57 mm at 250 µg/disc) was recorded with strains. The solvent methanol is known for its petroleum ether extract against Proteus vulgaris ability to isolate more antimicrobials and for fungi, the least zone of inhibition (7.0  0.50 from plants including tannins, polyphenols, mm) was observed against Candida tropicalis. terpenoids, saponins, xanthoxyllines, totarol, quassinoids, lactones, flavones and phenones, 4. Discussion while the water extracts could isolate only In the present study, different solvent viz., anthocyanins, starches, tannins, saponins, petroleum ether, chloroform, ethyl acetate and terpenoids, polypeptides and lectins. In general, methanol extracts of Psydrax dicoccos leaves were phenolic compounds possess specific physical, showed varied level of inhibitory activities against chemical and biological activities that make them the bacterial and fungal strains tested. All the useful as drugs. Phenolics were also responsible extracts of Psydrax dicoccos possessed significant for the antimicrobial, anti-inflammatory, anti- antibacterial and antifungal activity against all the feedant, anti-viral, anticancer and vasodilatory bacterial and fungal strains. The methanol leaf actions (Rievere, 2009). Phenolic compounds may

Umaiyambigai/ Indo – Asian Journal of Multidisciplinary Research (IAJMR), 2(1): 443 – 452 450 affect growth and metabolism of bacteria. They Finally, it can conclude that the methanol extract could have an activating or inhibiting effect on of Psydrax dicoccos can be used as a antimicrobial microbial growth according to their constitution substance for the treatment of microbial and concentration (Reguant 2000). Tannins were infections. used therapeutically as antiviral, antibacterial, Acknowledgement antiulcer and antioxidant agents. Many tannin containing drugs are used in the treatment of piles, The authors are thankful to Dr. K. inflammation, burns and as astringent (Kolodziej, Arumugam, Professor and Head, Department of 2005). Botany and authorities of Annamalai University for providing the necessary facilities. The MIC and MBC values of methanol extract were lower than that of other extracts of 6. Reference the Psydrax dicoccos showed potential 1) Ananatharaj, M., V. Venkatesalu, M. antibacterial effect against bacterial strains tested. Chandrasekaran and S. Sivasankari. 2004. Similar observations were made while studying Effect of seaweed liquid extract of Caulerpa the antimicrobial activity of seeds of Syzygium scalpelliformis on growth and biochemical jambolanum (Chandrasekaran, 2004), FAME constituents of black gram (Vigna mungo L.). extracts of certain marine macro algae (Ananatharaj Seaweed Res. Utilin., 26: 87 - 92. 2004) and leaves of Ipomoea pes-caprae (Chandrasekaran, 2005). The majority of plant 2) Ashik Mosaddik, M., K. E. Kabir and P. derived antimicrobial compounds generally have Hassan. 2000. Antibacterial activity of higher MICs than bacterial or fungal produced Alangium salviifolium flowers. Fitoterapia., antibiotics, thus limiting their therapeutic potential 71: 447 - 449. (Gibbons, 2004). 3) Avato, P., P. M. Vitali and A. Tava, 1997. In this study, the methanol extract of Antimicrobial activity of polyacetylenes leaves of Psydrax dicoccos showed the highest from Bellis perennis and synthetic derivatives. antibacterial activity against Gram positive Planta Medica, 63: 503 – 507. bacteria than Gram negative bacteria. The reason 4) Bauer, W. W., M. M Kirby, J. C. Scherris and for different sensitivity between Gram positive and M. Turck. 1966. Antibiotic susceptibility Gram negative bacteria could be ascribed to the testing by a standardized single disk method. morphological differences between these American J. Clin. Pathol., 45: 493 – 496. microorganisms (Avato et al., 1997). The Gram 5) Berkowitz, F. E. 1995. Antibiotic resistance in positive bacteria should be more susceptible since bacteria. Southern Med. Jour., 88: 797 – 804. they have only an outer peptidoglycan layer, which 6) Bhattacharjee, S. K. 1998 Handbook of is not an effective permeability barrier (Scherrer and Medicinal Plants. Pointer Pub, Jaipur-03, Gerhardt, 1971). The resistance of Gram India, pp. 1-6. negative bacteria towards antibacterial 7) Bonnet, R. 2004. Growing group of extended- substances was related to lipopolysaccharides in spectrum β-lactamases: the CTX-M enzymes. their outer membrane (Gao et al., 1999). Antimicrob. Agents Chemother., 48: 1–14. 5. Conclusion 8) Chandrasekaran, M and V. Venkatesalu. 2004. The antimicrobial activity of the Psydrax Antibacterial and antifungal activity of dicoccos methanol leaf extract, probably due to Syzygium jambolanum seeds. Journal of the recognized phytoconstituents, further confirms Ethanopharmacology, 91: 105 - 108. its use as a health cure in traditional medicine. 9) Chandrasekaran, M., V. Venkatesalu, S. Bioactive substances from this plant as a result be Ananatharaj and M. S. Sivasankari. 2005. employed in the formulation of antifungal agents Antibacterial activity of fatty acid methyl for the healing of a variety of microbial infections.

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